|
1. Target Preparation: Using the starting sample of total RNA as a template, double stranded cDNA is synthesized, and from that, biotin-labeled cRNA is synthesized. The labeled cRNA is then fragmented.
2. Target Hybridization: A hybridization cocktail is prepared using the fragmented cRNA, probe array controls, BSA, and Herring Sperm DNA. The cocktail is then hybridized onto the Genechip for 16 hours.
3. Experiment Setup and Fluidics: After hybridization is complete, specified experiment information is defined in the operating software, GCOS (Gene Chip Operating Software). The chips are washed and stained using the fluidics station.
4. Probe Array Scan and Data Output: The hybridized chip is scanned using the GeneChip scanner, which outputs a data image file (.dat). The system then computes a cell intensity file (.cel) which averages the intensity of each individual cell on the array. The data collected is then used to form the analysis file (.chp) that contains all of the data output in text format. All data files, along with an excel file of the analysis, is placed on a CD and returned to the client. |